SN 1544-2005進(jìn)出口動(dòng)物源性食品中玉米赤霉醇?xì)埩袅康臋z驗(yàn)方法 高效液相色譜-質(zhì)譜質(zhì)譜法

中華人民共和國(guó)出入境檢驗(yàn)檢疫行業(yè)標(biāo)準(zhǔn)進(jìn)出口動(dòng)物源性食品中玉米赤霉醇?xì)埩袅康臋z驗(yàn)方法高效液相色譜-質(zhì)譜/質(zhì)譜法2005-02-17發(fā)布國(guó)家質(zhì)量監(jiān)督檢驗(yàn)檢疫總局I本標(biāo)準(zhǔn)的附錄A為資料性附錄。本標(biāo)準(zhǔn)由國(guó)家認(rèn)證認(rèn)可監(jiān)督管理委員會(huì)提出并歸口。本標(biāo)準(zhǔn)系首次發(fā)布的出入境檢驗(yàn)檢疫行業(yè)標(biāo)準(zhǔn)。1進(jìn)出口動(dòng)物源性食品中玉米赤霉醇?xì)埩袅勘緲?biāo)準(zhǔn)規(guī)定了動(dòng)物源性食品中玉米赤霉醇?xì)埩袅繖z驗(yàn)的抽樣、制樣和高效液相色譜-質(zhì)譜/質(zhì)譜檢以不超過2500件為一檢驗(yàn)批。抽樣數(shù)量見表1。表1單位為件最低抽樣數(shù)126～1005101～250251～500501～10001001～25002.3抽樣方法按2.2規(guī)定的抽樣件數(shù)隨機(jī)抽取，逐件開啟。從每件中至少取一袋作為原始樣品。如每件中無小包裝或有小包裝，但每袋小包裝質(zhì)量超過2kg,則可用滅菌刀具從每件中割取不少于500g,混合后置于清潔容器內(nèi)作為混合原始樣品。混合原始樣品的總中質(zhì)量不少于2kg,加封后，標(biāo)明標(biāo)記，及時(shí)送交從混合原始樣品中取出部分有代表性的樣品，充分混勻，用四分法縮分至不少于500g,均分成兩3.1方法提要?jiǎng)驖{組織樣品經(jīng)葡萄糖苷酸酶水解后，依次用乙醚和氫氧化鈉溶液提取玉米赤霉醇。氫氧化鈉提取液中和后經(jīng)C??固相萃取柱凈化。甲醇洗脫液經(jīng)蒸發(fā)，定容后供高效液相色譜-質(zhì)譜/質(zhì)譜儀測(cè)定，外標(biāo)法定量。23.2.7磷酸：純度>85%;密度1.7g/mL。3.2.8葡萄糖苷酸酶。酸調(diào)節(jié)至pH4.8。制成0.10mg/mL的標(biāo)準(zhǔn)儲(chǔ)備液。儲(chǔ)備液保存于-18℃。稱取5g試樣(精確至0.01g),置于潔凈的50mL具塞離心管中，加入10mL乙酸鈉緩沖溶液(3.2.13)和25μL葡萄糖苷酸酶，旋渦混勻，于37℃中保溫8h～14h。34玉米赤霉醇添加濃度及其回收率數(shù)據(jù)：——1pg/kg時(shí)，回收率72%;——5μg/kg時(shí)，回收率74%;5SN/T1544—2005(資料性附錄)0ThisstandardwasproposedbyandisunderthechargeofNationRegulatoryCommissionforCertifi-ThisstandardwasdraftedbyShanghaiEntry-ExitinspectionandQuarantineBureauofthePeople'sRepublicofChina.ThisstandardwasmainlydraftedbyFangXiaoming,ChenJiahua,NiXinluandTangYifeng.Thisstandardispromulgatedforthefirsttime.7productsforimportandexport—HPLC-MS/MSThisstandardspecifianimaloriginalproductsbyhigh-performanceliquidchromatography-tandemmassandspecification,shouldbTable1Minimumnumberofpackagestobetaken126～1005101～250251~500501～10001001～2500samplewasdevidedintwoandhomogenized.Packegeandlabel.8Thetestsampleshouldbestoredat-18℃.AhomogenizedtissuesampleishydrolyzedwithB-glucuronidaseandthehydrolysateisextractedwithethylether.Thesupernatantisevaporatedtodrynessandtheresidueisdissolvedinchloroformandre-extractedwithsodiumhydroxidesoCasolid-phaseextractioncartridge,andanalyzedbyelectrosprayHPLC-MS/MSinnegativeionmode.Reagentswereofanalytical-reagentgradeexceptforthespecificreagents.Thewaterusedwasdoublydistilledordeionizedw3.2.1Acetonitrile:LCgrade.3.2.6Anhydroussodiumacetate.3.2.7Phosphoricacid:purity>85%;3.2.9Zeranol(a-zearalanol):purity97%.withwater.3.2.13Sodiumacetatebuffer,50mmol/L:Take4.1gofanhydroussodiumacetate,dissolveand9diluteto1000mLwithwater.AdjustpHto4.8withglacialaceticacid.3.2.14Zeranolstocksolution,0.10mg/mL:Accuratelyweighproperzeranosolvedwithacetonitrile.Tstocksolution(3.2.14)into100mLvolumetrictively.ThesolutionsanolIntermediatesolutions(3.2.15).Thematrix-matchedstandardsarestoredat-week.3.2.17Cisolid-phaseextractioncartridge,500m3.3.1High-performanceliquidchromatography-electrospraytandemmassspectrometry.3.3.3VacuummanifoldforsolidWeigh5.0g±0.1gofhomogenizedtissuesampleintoa50mLcentrifugetube.Add10mLofsodi-SN/T1544—2005umacetatebuffer(3.2.13)and25μLofβ-glucuronidase,mixwell,andthenincubat(3.2.13)and2mLofmethan200pLofacetonitrileforanalb)Mobilephase:acetonitrile-water(40+60)containing0.025%aceticacid.d)Columntemperature:22℃.a)lonizationmode:negativeelectrospray.b)Conevoltage:50V.c)Capillaryvoltage:3kV.d)Collisionvoltage:20V.e)Analyzervacuum:<3mPa(3×10~5mbar).f)Collisiongas:argon.h)Nebulisergasflow:480L/h.i)Multiplereactionmonitoring(MRM)mode:m/z321→303,m/z321→277.m/forAnalyzematrix-matchedstandards(3.2.16)inorderofincreasingconcentration,obtainingthema-withoutadditionofthesample.Theconcentrationofzeranolinsampleiscalculatedwithchromatographicdataprocessoro…………(1)Where:X—theconcentrationofanalyteofinterestinsample,ug/kg:c~—theconcentrationofanalytefromcalibrationcurve,ng/mL;V--—finalvolumeofsampleextract,mL;m——weightofsample,g.Note:Theblankvalueshouldbesubtracte